Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 546/羊抗鼠IgG (H+L)交叉吸附荧光二抗,Alexa Fluor 546

货号：A-11003

规格：500 µL

价格：3191

产品类型：荧光二抗

品牌：Thermo Fisher

抗原：Gamma Immunoglobins Heavy and Light chains

物种：小鼠

宿主：山羊

抗体亚型：IgG

荧光染料：Alexa Fluor 546

点击查看所有Alexa Fluor 荧光二抗

抗体类型:	荧光二抗	同型对照：
浓度：	2mg/mL	用法：	4µg/mL(ICC);4µg/mL（IF);1-10µg/mL（Flow)

产品详细信息To minimize cross-reactivity, these goat anti-mouse IgG (H+L) whole secondary antibodies have been affinity purified and cross-adsorbed against human IgG and human serum prior to conjugation. Cross-adsorption or pre-adsorption is a purification step to increase specificity of the antibody resulting in higher sensitivity and less background staining. The secondary antibody solution is passed through a column matrix containing immobilized serum proteins from potentially cross-reactive species. Only the nonspecific-binding secondary antibodies are captured in the column, and the highly specific secondaries flow through. The benefits of this extra step are apparent in multiplexing/multicolor-staining experiments (e.g., flow cytometry) where there is potential cross-reactivity with other primary antibodies or in tissue/cell fluorescent staining experiments where there are may be the presence of endogenous immunoglobulins.Alexa Fluor dyes are among the most trusted fluorescent dyes available today. Invitrogen™ Alexa Fluor 546 dye is a bright, orange-fluorescent dye with excitation ideally suited to the 546 nm laser line. For stable signal generation in imaging and flow cytometry, Alexa Fluor 546 dye is pH-insensitive over a wide molar range. Probes with high fluorescence quantum yield and high photostability allow detection of low-abundance biological structures with great sensitivity. Alexa Fluor 546 dye molecules can be attached to proteins at high molar ratios without significant self-quenching, enabling brighter conjugates and more sensitive detection. The degree of labeling for each conjugate is typically 2-8 fluorophore molecules per IgG molecule; the exact degree of labeling is indicated on the certificate of analysis for each product lot.Using conjugate solutions: Centrifuge the protein conjugate solution briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step will help eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining. Because staining protocols vary with application, the appropriate dilution of antibody should be determined empirically. For the fluorophore-labeled antibodies a final concentration of 1-10 µg/mL should be satisfactory for most immunohistochemistry and flow cytometry applications.靶标信息We offer an extensive line of Invitrogen™ secondary antibody conjugates with well-characterized specificity and labeled with a wide selection of premium fluorescent dyes, including Invitrogen™ Alexa Fluor™ fluorescent dyes. Fluorescent secondary antibody conjugates are useful in the detection, sorting, or purification of its specified target and ideal for fluorescence microscopy and confocal laser scanning microscopy, flow cytometry, and fluorescent western detection. The breadth of fluorescent markers we offer allows our reagents to be tailored to almost any fluorescent detection system.Secondary antibodies may be provided in three formats: whole IgG, divalent F(ab')2 fragments, and monovalent Fab fragments. Because of the high degree of conservation in the structure of many immunoglobulin domains, most class-specific secondary antibodies must be affinity-purified and cross-adsorbed to achieve minimal cross-reaction with other immunoglobulins.Our secondary antibody conjugates are most commonly prepared by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (e.g., immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents. In the first round of purification, whole immunoglobulins binding to the immunizing antibody are recovered and mainly consist of the ~150-kDa IgG class. Further purification, for example, with Protein A or G, removes all unwanted immunoglobulin classes except the affinity-purified antibodies that react with the target-specific immunoglobulin heavy and/or light chains.For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

数据

Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11003) in IFImmunofluorescence analysis of Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 546 (Product # A-11003) was performed using HeLa cells stained with alpha Tubulin (236-10501) Mouse Monoclonal Antibody (Product # A11126). The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, blocked with 1% BSA for 1 hour and labeled with 2 µg/mL of mouse primary antibody for 3 hours at room temperature. Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 546 (Product # A-11003) was used at concentration of 4 µg/mL in phosphate buffered saline containing 0.2 % BSA for 45 minutes at room temperature, for detection of alpha Tubulin in the cytoplasm (Panel a: red). Nuclei (Panel b: blue) were stained with DAPI in SlowFade® Gold Antifade Mountant (Product # S36938). F-actin was stained with Alexa Fluor® 488 Phalloidin (Product # A12379, 1:300) (Panel c: green). Panel d represents the composite image. No nonspecific staining was observed with the secondary antibody alone (panel f), or with an isotype control (panel e). The images were captured at 60X magnification. Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11003) in IFComparison of the photobleaching rates of the Alexa Fluor® 488 and Alexa Fluor® 546 dyes and the well-known fluorescein and Cy3 fluorophores. The cytoskeleton of bovine pulmonary artery endothelial cells (BPAEC) was labeled with (top series) Alexa Fluor® 488 phalloidin (Product # A12379) and mouse monoclonal anti-alpha-tubulin antibody (Product # A11126) in combination with Alexa Fluor® 546 goat anti-mouse IgG antibody (Product # A-11003) or (bottom series) fluorescein phalloidin (F432) and the anti-alpha-tubulin antibody in combination with a commercially available Cy3 goat anti-mouse IgG antibody. The pseudocolored images were taken at 30-second intervals (0, 30, 90, and 210 seconds of exposure from left to right). The images were acquired with bandpass filter sets appropriate for fluorescein and rhodamine. Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11003) in IFCultured axolotl sensory neurons stained with our anti-human neuronal protein HuC/HuD antibody (anti-Hu, Product # A-21271) and detected using the red-orange-fluorescent Alexa Fluor® 546 goat anti-mouse IgG antibody (Product # A-11003). Actin was labeled using green-fluorescent Alexa Fluor® 488 phalloidin (Product # A12379) and nuclei were stained with blue-fluorescent Hoechst 33342 (Product # H1399, H3570, H21492). The image was contributed by Josh Gross and Linda Barlow, University of Denver, Denver, Colorado. Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody (A-11003) in IFPhotostability comparison of Alexa Fluor® 488 phalloidin (top panel, Product # A12379) and fluorescein phalloidin (bottom panel, Product # F432). Bovine pulmonary artery endothelial cells labeled with 5 units/mL of Alexa Fluor® 488 phalloidin or fluorescein phalloidin, and 10 µg/mL of Alexa Fluor® 546 goat anti-mouse IgG (red-fluorescent tubulin staining, Product # A-11003). Images were acquired every three seconds for a total of 50 seconds using a Quantix camera (Photometrics).

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参考文献：

1.Journal of cellular physiologyMactosylceramide prevents glial cell overgrowth by inhibiting insulin and fibroblast growth factor receptor signaling.Loss of O-GlcNAc glycosylation in forebrain excitatory neurons induces neurodegeneration.3.Proceedings of the National Academy of Sciences of the United States of AmericaSpemann organizer gene Goosecoid promotes delamination of neuroblasts from the otic vesicle.4.Molecular visionmice.5.Frontiers in neuroanatomyAdeno-Associated Viral Vectors Serotype 8 for Cell-Specific Delivery of Therapeutic Genes in the Central Nervous System.

技术参数

产品应用 ICC;IF;Flow

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